Full-field fluorescence lifetime dual-comb microscopy using spectral mapping and frequency multiplexing of dual-comb optical beats
| Autor: | Takeshi Yasui, Yu Tokizane, Takeo Minamikawa, Hidenori Koresawa, Takahiko Mizuno, Eiji Hase, Hirotsugu Yamamoto, Ryo Oe |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Fluorescence-lifetime imaging microscopy
Multidisciplinary Materials science Pixel business.industry Phase (waves) SciAdv r-articles Optics 02 engineering and technology 021001 nanoscience & nanotechnology 01 natural sciences Fluorescence Photobleaching Frequency-division multiplexing 010309 optics 0103 physical sciences Microscopy Radio frequency 0210 nano-technology business Research Articles Research Article Applied Physics |
| Zdroj: | Science Advances |
| ISSN: | 2375-2548 |
| DOI: | 10.1126/sciadv.abd2102 |
| Popis: | Use of dual-comb optical beats enables scan-less FLIM via one-to-one correspondence between 2D image pixels and RF beat signals. Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for quantitative fluorescence imaging because fluorescence lifetime is independent of concentration of fluorescent molecules or excitation/detection efficiency and is robust to photobleaching. However, since most FLIMs are based on point-to-point measurements, mechanical scanning of a focal spot is needed for forming an image, which hampers rapid imaging. Here, we demonstrate scan-less full-field FLIM based on a one-to-one correspondence between two-dimensional (2D) image pixels and frequency-multiplexed radio frequency (RF) signals. A vast number of dual-comb optical beats between dual optical frequency combs are effectively adopted for 2D spectral mapping and high-density frequency multiplexing in the RF region. Bimodal images of fluorescence amplitude and lifetime are obtained with high quantitativeness from amplitude and phase spectra of fluorescence RF comb modes without the need for mechanical scanning. The parallelized FLIM will be useful for rapid quantitative fluorescence imaging in life science. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|